Bovine Dendritic Cell Growth Kit
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Bovine Dendritic Cell Growth Kit
- Product Type
- Bovine Dendritic Cell Growth Kit
|Bovine dendritic cell growth kit contains a cocktail of biologically active interleukin-4 (IL-4) and granulocyte/macrophage-colony stimulating factor (GM-CSF) that have been premixed at optimal concentrations to induce dendritic cell development from peripheral blood-derived bovine (cattle) monocytes.|
- Target Species
- Product Form
- Mixed recombinant bovine Interleukin-4 and bovine GM-CSF – supplied as a liquid
- Recombinant cytokines expressed in mammalian Chinese Hamster Ovary (CHO) cells using the pEE14® vector grown in antibiotic free media and USDA-approved dialysed FCS which has been screened for BVDV and virus growth by PCR.
- Preservative Stabilisers
- None present
- Endotoxin Level
- < 0.5 EU/ml
- For research purposes only
- 6 months from date of despatch
- This reagent was produced as part of the BBSRC/SEERAD Immunological Toolbox. The kit development was also supported by the European Community’s Seventh Framework Programme (FP7, 2007-2013), Research Infrastructures action, under the grant agreement No. FP7-228394 (NADIR)
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature this recombinant protein. Should this product contain a precipitate we recommend microcentrifugation before use.
|Application Name||Verified||Min Dilution||Max Dilution|
- Instructions For Use
- 1. Prepare peripheral blood mononuclear cells (PBMC) from heparinised blood by density gradient centrifugation.
2. Purify CD14+ve cells by labelling PBMC with CD14 mAb and utilise magnetic bead or flow cytometric separation techniques.
3. Resuspend the isolated CD14+ve cells at a concentration of 1x106 cells/ml in tissue culture medium (TCM = RPMI or equivalent + 10% foetal calf serum) containing a final dilution of 1:20 of PBP015KZZ .
4. Add 3ml of cell suspension to each well of a 6 well tissue culture plate.
5. Culture cells in a humidified atmosphere of 5% CO2 in air, at approximately 37oC.
6. Culture cells for 3 days. The cells may then be harvested and used for other procedures including immunophenotyping (as required).
7. If a longer culture period is required the cells must be ‘fed’ with new TCM containing cytokines on day 3:
Carefully remove 1ml spent medium from each well, care is required to avoid disturbing the cells.
Add 1.5ml fresh, pre-warmed TCM containing cytokines at 1:20 to each well and re-culture the DC for required culture period (typically up to 7 days).
8. At the end of the culture period adherent and non-adherent cells can be pooled for use in immunoassays and phenotyped (as required). Adherent cells may require a dissociation step to remove them from the plate.
References for Bovine Dendritic Cell Growth Kit
Walters, A.A. et al. (2015) Assessment of the enhancement of PLGA nanoparticle uptake by dendritic cells through the addition of natural receptor ligands and monoclonal antibody.
Vaccine. 33 (48): 6588-95.
Werling, D. et al. (1999) Involvement of caveolae in the uptake of respiratory syncytial virus antigen by dendritic cells.
J Leukoc Biol. 66 (1): 50-8.
Hope, J.C. et al. (2000) Dendritic cells induce CD4+ and CD8+ T-cell responses to Mycobacterium bovis and M. avium antigens in Bacille Calmette Guérin vaccinated and nonvaccinated cattle.
Scand J Immunol. 52 (3): 285-91.
Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"View all Anti-Bovine Products
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